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At least 1,117 records · Page 62

Data for High-Level β-Carotene Production from Xylose by Engineered Saccharomyces cerevisiae without Overexpression of a Truncated HMG1 (t HMG1 )

β‐Carotene is a natural pigment and health‐promoting metabolite, and has been widely used in the nutraceutical, feed, and cosmetic industries. Here, we engineered a GRAS yeast Saccharomyces cerevisiae to produce β‐carotene from xylose, the second most abundant and inedible sugar component of lignocellulose biomass. Specifically, a β‐carotene biosynthetic pathway containing crtYB, crtI, and crtE from Xanthophyllomyces dendrorhous was introduced into a xylose‐fermenting S. cerevisiae . The resulting strain produced β‐carotene from xylose at a titer threefold higher than from glucose. Interestingly, overexpression of t HMG1 , which has been reported as a critical genetic perturbation to enhance metabolic fluxes in the mevalonate pathway and β‐carotene production in yeast when glucose is used, did not further improve the production of β‐carotene from xylose. Through fermentation profiling, metabolites analysis, and transcriptional studies, we found the advantages of using xylose as a carbon source, instead of glucose, for β‐carotene production to be a more respiratory feature of xylose consumption, a larger cytosolic acetyl‐CoA pool, and an upregulated expression level of rate‐limiting genes in the β‐carotene‐producing pathway, including ACS1 and HMG1 . As a result, 772.8 mg/L of β‐carotene was obtained in a fed‐batch bioreactor culture with xylose feeding. Considering the inevitable large scale production of xylose when cellulosic biomass‐based bioeconomy is implemented, our results suggest xylose utilization is a promising strategy for overproduction of carotenoids and other isoprenoids in engineered S. cerevisiae .

Conversion↗

Microfabrication of a Chamber for High-Resolution, In Situ Imaging of the Whole Root for Plant–Microbe Interactions

Fabricated ecosystems (EcoFABs) offer an innovative approach to in situ examination of microbial establishment patterns around plant roots using nondestructive, high-resolution microscopy. Previously high-resolution imaging was challenging because the roots were not constrained to a fixed distance from the objective. Here, we describe a new ‘Imaging EcoFAB’ and the use of this device to image the entire root system of growing Brachypodium distachyon at high resolutions (20×, 40×) over a 3-week period. The device is capable of investigating root–microbe interactions of multimember communities. We examined nine strains of Pseudomonas simiae with different fluorescent constructs to B. distachyon and individual cells on root hairs were visible. Succession in the rhizosphere using two different strains of P. simiae was examined, where the second addition was shown to be able to establish in the root tissue. The device was suitable for imaging with different solid media at high magnification, allowing for the imaging of fungal establishment in the rhizosphere. Overall, the Imaging EcoFAB could improve our ability to investigate the spatiotemporal dynamics of the rhizosphere, including studies of fluorescently-tagged, multimember, synthetic communities.

59 BASIC BIOLOGICAL SCIENCES↗

Influence of heat treatment on mechanical properties of 300M Steel

The plane strain fracture toughness and tensile strength response of 300M Steel to a wide variety of austenitizing and tempering temperatures were investigated. The results make it possible for one to select heat treatments which provide an optimum combination of strength and toughness for a variety of structural applications. In particular, improvements in toughness on the order of 20% were found possible with no loss in tensile properties by increasing the austenitizing temperature from the currently employed 1144 K to 1255 K or higher, and this change in heat treatment therefore appears worthy of general implementation.

Youngblood, J. L.↗

Superplastic forming of ceramic insulation

Superplasticity has been demonstrated in many fine-grained structural ceramics and ceramic composites, including yttria-stabilized tetragonal zirconia polycrystal (YTZP), alumina, and Al2O3-reinforced zirconia (Al2O3/YTZ) duplex composites and SiC-reinforced Si3N4. These superplastic ceramics obviously offer the potential benefit of forming net shape or near net shape parts. This could be particularly useful for forming complicated shapes that are difficult to achieve using conventional forming techniques, or require elaborate, subsequent machining. In the present study, we successfully demonstrated the following: (1) superplastic 3Y-TXP and 20 percent Al2O3/YTZ composite have for the first time been successfully deformed into hemispherical caps via a biaxial gas-pressure forming technique; (2) no experimental difficulty was encountered in applying the required gas pressures and temperatures to achieve the results, thus, it is certain that higher rates of deformation than those presented in this study will be possible by using the current test apparatus at higher temperatures and pressures; and (3) an analytical model incorporating material parameters, such as variations during forming in the strain rate sensitivity exponent and grain growth-induced strain hardening, is needed to model accurately and therefore precisely control the biaxial gas-pressure forming of superplastic ceramics. Based on the results of this study, we propose to fabricate zirconia insulation tubes by superplastic extrusion of zirconia polycrystal. This would not only reduce the cost, but also improve the reliability of the tube products.

Nieh, T. G.↗

Multifunctional Nanotube Polymer Nanocomposites for Aerospace Applications: Adhesion between SWCNT and Polymer Matrix

Multifunctional structural materials can enable a novel design space for advanced aerospace structures. A promising route to multifunctionality is the use of nanotubes possessing the desired combination of properties to enhance the characteristics of structural polymers. Recent nanotube-polymer nanocomposite studies have revealed that these materials have the potential to provide structural integrity as well as sensing and/or actuation capabilities. Judicious selection or modification of the polymer matrix to promote donor acceptor and/or dispersion interactions can improve adhesion at the interface between the nanotubes and the polymer matrix significantly. The effect of nanotube incorporation on the modulus and toughness of the polymer matrix will be presented. Very small loadings of single wall nanotubes in a polyimide matrix yield an effective sensor material that responds to strain, stress, pressure, and temperature. These materials also exhibit significant actuation in response to applied electric fields. The objective of this work is to demonstrate that physical properties of multifunctional material systems can be tailored for specific applications by controlling nanotube treatment (different types of nanotubes), concentration, and degree of alignment.

Park, Cheol↗

Understanding Earthquake Fault Systems Using QuakeSim Analysis and Data Assimilation Tools

We are using the QuakeSim environment to model interacting fault systems. One goal of QuakeSim is to prepare for the large volumes of data that spaceborne missions such as DESDynI will produce. QuakeSim has the ability to ingest distributed heterogenous data in the form of InSAR, GPS, seismicity, and fault data into various earthquake modeling applications, automating the analysis when possible. Virtual California simulates interacting faults in California. We can compare output from long time history Virtual California runs with the current state of strain and the strain history in California. In addition to spaceborne data we will begin assimilating data from UAVSAR airborne flights over the San Francisco Bay Area, the Transverse Ranges, and the Salton Trough. Results of the models are important for understanding future earthquake risk and for providing decision support following earthquakes. Improved models require this sensor web of different data sources, and a modeling environment for understanding the combined data.

earthquake simulations↗

Verification and Validation of a Three-Dimensional Generalized Composite Material Model

A general purpose orthotropic elasto-plastic computational constitutive material model has been developed to improve predictions of the response of composites subjected to high velocity impact. The three-dimensional orthotropic elasto-plastic composite material model is being implemented initially for solid elements in LS-DYNA as MAT213. In order to accurately represent the response of a composite, experimental stress-strain curves are utilized as input, allowing for a more general material model that can be used on a variety of composite applications. The theoretical details are discussed in a companion paper. This paper documents the implementation, verification and qualitative validation of the material model using the T800-F3900 fiber/resin composite material

Plasticity↗

Verification and Validation of a Three-Dimensional Generalized Composite Material Model

A general purpose orthotropic elasto-plastic computational constitutive material model has been developed to improve predictions of the response of composites subjected to high velocity impact. The three-dimensional orthotropic elasto-plastic composite material model is being implemented initially for solid elements in LS-DYNA as MAT213. In order to accurately represent the response of a composite, experimental stress-strain curves are utilized as input, allowing for a more general material model that can be used on a variety of composite applications. The theoretical details are discussed in a companion paper. This paper documents the implementation, verification and qualitative validation of the material model using the T800- F3900 fiber/resin composite material.

Finite Element Method↗

Impact and Penetration of Thin Aluminum 2024 Flat Panels at Oblique Angles of Incidence

The U.S. Federal Aviation Administration (FAA) and the National Aeronautics and Space Administration (NASA) are actively involved in improving the predictive capabilities of transient finite element computational methods for application to safety issues involving unintended impacts on aircraft and aircraft engine structures. One aspect of this work involves the development of an improved deformation and failure model for metallic materials, known as the Tabulated Johnson-Cook model, or MAT224, which has been implemented in the LS-DYNA commercial transient finite element analysis code (LSTC Corp., Livermore, CA) (Ref. 1). In this model the yield stress is a function of strain, strain rate and temperature and the plastic failure strain is a function of the state of stress, temperature and strain rate. The failure criterion is based on the accumulation of plastic strain in an element. The model also incorporates a regularization scheme to account for the dependency of plastic failure strain on mesh size. For a given material the model requires a significant amount of testing to determine the yield stress and failure strain as a function of the three-dimensional state of stress, strain rate and temperature. In addition, experiments are required to validate the model. Currently the model has been developed for Aluminum 2024 and validated against a series of ballistic impact tests on flat plates of various thicknesses (Refs. 1 to 3). Full development of the model for Titanium 6Al-4V is being completed, and mechanical testing for Inconel 718 has begun. The validation testing for the models involves ballistic impact tests using cylindrical projectiles impacting flat plates at a normal incidence (Ref. 2). By varying the thickness of the plates, different stress states and resulting failure modes are induced, providing a range of conditions over which the model can be validated. The objective of the study reported here was to provide experimental data to evaluate the model under more extreme conditions, using a projectile with a more complex shape and sharp contacts, impacting flat panels at oblique angles of incidence.

Ruggeri, Charles R.↗

Signal Origin of Electrochemical Strain Microscopy and Link to Local Chemical Distribution in Solid State Electrolytes

Electrochemical strain microscopy (ESM) is a distinguished method to characterize Li-ion mobility in energy materials with extremely high spatial resolution. The exact origin of the cantilever deflection when the technique is applied on solid state electrolytes (SSEs) is currently discussed in the literature. Under-standing local properties and influences on ion mobility in SSEs is of utmost importance to improve such materials for next generation batteries. Here, the exact signal formation process of ESM when applied on sodium super ionic conductor (NASICON)-type SSE containing Na- and Li-ions is investigated. Changes in the dielectric properties, which are linked to the local chemical composition, are found to be responsible for the observed contrast in the deflection of the cantilever instead of a physical volume change as a result of Vegard´s Law. The cantilever response is strongly reduced in areas of high sodium content which is attributed to a reduction of the tip-sample capacitance in comparison to areas with high lithium content. This is the first time a direct link between electrostatic forces in contact mode and local chemical information is demonstrated on SSEs. Furthermore, the results open up new possibilities in information gain since dielectric proper-ties are sensitive to subtle changes in local chemical composition.

36 MATERIALS SCIENCE↗

Development of a Genome-Scale Metabolic Model of Clostridium thermocellum and Its Applications for Integration of Multi-Omics Datasets and Computational Strain Design

Solving environmental and social challenges such as climate change requires a shift from our current non-renewable manufacturing model to a sustainable bioeconomy. To lower carbon emissions in the production of fuels and chemicals, plant biomass feedstocks can replace petroleum using microorganisms as biocatalysts. The anaerobic thermophile Clostridium thermocellum is a promising bacterium for bioconversion due to its capability to efficiently degrade lignocellulosic biomass. However, the complex metabolism of C. thermocellum is not fully understood, hindering metabolic engineering to achieve high titers, rates, and yields of targeted molecules. In this study, we developed an updated genome-scale metabolic model of C. thermocellum that accounts for recent metabolic findings, has improved prediction accuracy, and is standard-conformant to ensure easy reproducibility. We illustrated two applications of the developed model. We first formulated a multi-omics integration protocol and used it to understand redox metabolism and potential bottlenecks in biofuel (e.g., ethanol) production in C. thermocellum. Second, we used the metabolic model to design modular cells for efficient production of alcohols and esters with broad applications as flavors, fragrances, solvents, and fuels. The proposed designs not only feature intuitive push-and-pull metabolic engineering strategies, but also present novel manipulations around important central metabolic branch-points. We anticipate the developed genome-scale metabolic model will provide a useful tool for system analysis of C. thermocellum metabolism to fundamentally understand its physiology and guide metabolic engineering strategies to rapidly generate modular production strains for effective biosynthesis of biofuels and biochemicals from lignocellulosic biomass.

59 BASIC BIOLOGICAL SCIENCES↗

High–level β–carotene production from xylose by engineered Saccharomyces cerevisiae without overexpression of a truncated HMG1 (t HMG1 )

Beta-carotene is a natural pigment and health-promoting metabolite, and has been widely used in the nutraceutical, feed, and cosmetic industries. Here, we engineered a GRAS yeast Saccharomyces cerevisiae to produce beta-carotene from xylose, the sec- ond most abundant and inedible sugar component of lignocellulose biomass. Specifically, a beta-carotene biosynthetic pathway containing crtYB, crtI, and crtE from Xanthophyllomyces dendrorhous was introduced into a xylose-fermenting S. cerevisiae. Here, the resulting strain produced beta-carotene from xylose at a titer threefold higher than from glucose. Interestingly, overexpression of tHMG1, which has been reported as a critical genetic perturbation to enhance metabolic fluxes in the mevalonate pathway and beta-carotene production in yeast when glucose is used, did not further improve the production of beta-carotene from xylose. Through fermentation profiling, metabolites analysis, and transcriptional studies, we found the advantages of using xylose as a carbon source, instead of glucose, for beta-carotene production to be a more respiratory feature of xylose consumption, a larger cytosolic acetyl!CoA pool, and an upregulated expression level of rate-limiting genes in the beta-carotene!producing pathway, including ACS1 and HMG1. As a result, 772.8 mg/L of beta-carotene was ob- tained in a fed-batch bioreactor culture with xylose feeding. Considering the in- evitable large scale production of xylose when cellulosic biomass!based bioeconomy is implemented, our results suggest xylose utilization is a promising strategy for overproduction of carotenoids and other isoprenoids in engineered S. cerevisiae.

09 BIOMASS FUELS↗

Significant disparity of non-basal dislocation activities in hot-rolled highly-textured Mg and Mg-3Al-1Zn alloy under tension

It is well-established that Mg-3Al-1Zn (AZ31) alloy exhibits much better tensile ductility than pure Mg. However, the underlying mechanisms for such difference still remain relatively unexplored at the dislocation level. In this work, we deformed hot-rolled highly-textured pure Mg and AZ31 samples with similar initial microstructures (i.e., grain size and texture) under tension along the rolling direction. Apparent shear banding was only observed in the pure Mg samples from the early stage of the deformation. Shear bands could act as easy paths for crack propagation, leading to low ductility in pure Mg. Apparent shear banding was not noted in deformed AZ31 samples. We then investigated the deformation mechanisms at the dislocation level using transmission electron microscopy. Systematic tilting experiments and statistical analyses of multiple grains at different strain levels revealed a significant disparity of non-basal dislocation activities between pure Mg and AZ31. For pure Mg, dislocations were activated since the early stage of plastic deformation. For AZ31, dislocations were mostly absent at all strain levels, even in the strain-to-failure samples. Non-basal dislocations, including prismatic and pyramidal dislocations, were observed. The promotion of the non-basal dislocation activities and the suppression of dislocations in AZ31 are expected to offer more sustainable hardening, which could elucidate the absence of apparent shear banding and much-improved ductility in AZ31 compared to pure Mg.

Zhao, Dexin↗

New self-consistent homogenization for thermo-elastic polycrystals with imperfect interfaces

Here we present a new thermo-elastic self-consistent formulation to calculate the thermo-mechanical response of polycrystalline aggregates with imperfect interfaces. The new model is based on an improved treatment of the effect of imperfect interfaces on the micromechanical response of the grains compared to pre-existing models. New expressions/algorithms are derived for the effective elastic and thermal properties, localization relations, and calculation of intragranular field fluctuations. In addition, derivation of a novel integral equation for the strain field in a heterogeneous elastic medium subjected to arbitrary thermal strain field and displacement jumps is presented. The new model is applied to study the effective thermo-elastic properties and average field fluctuations as a function of the interface properties for imperfectly-bonded triaminotrinitrobenzene (TATB) polycrystalline granular aggregates. The proposed formulation is general in the sense that it can be extended to other material systems and deformation regimes that require a proper consideration of the effect of imperfect interface properties on the material's thermo-mechanical response.

36 MATERIALS SCIENCE↗

Operando neutron diffraction reveals mechanisms for controlled strain evolution in 3D printing

Abstract Residual stresses affect the performance and reliability of most manufactured goods and are prevalent in casting, welding, and additive manufacturing (AM, 3D printing). Residual stresses are associated with plastic strain gradients accrued due to transient thermal stress. Complex thermal conditions in AM produce similarly complex residual stress patterns. However, measuring real-time effects of processing on stress evolution is not possible with conventional techniques. Here we use operando neutron diffraction to characterize transient phase transformations and lattice strain evolution during AM of a low-temperature transformation steel. Combining diffraction, infrared and simulation data reveals that elastic and plastic strain distributions are controlled by motion of the face-centered cubic and body-centered cubic phase boundary. Our results provide a new pathway to design residual stress states and property distributions within additively manufactured components. These findings will enable control of residual stress distributions for advantages such as improved fatigue life or resistance to stress-corrosion cracking.

36 MATERIALS SCIENCE↗

Rapid mini‐chromosome divergence among fungal isolates causing wheat blast outbreaks in Bangladesh and Zambia

Summary The fungal pathogen,Magnaporthe oryzae Triticumpathotype, causing wheat blast disease was first identified in South America and recently spread across continents to South Asia and Africa. Here, we studied the genetic relationship among isolates found on the three continents. Magnaporthe oryzaestrains closely related to a South American field isolate B71 were found to have caused the wheat blast outbreaks in South Asia and Africa. Genomic variation among isolates from the three continents was examined using an improved B71 reference genome and whole‐genome sequences. We found strong evidence to support that the outbreaks in Bangladesh and Zambia were caused by the introductions of genetically separated isolates, although they were all close to B71 and, therefore, collectively referred to as the B71 branch. In addition, B71 branch strains carried at least one supernumerary mini‐chromosome. Genome assembly of a Zambian strain revealed that its mini‐chromosome was similar to the B71 mini‐chromosome but with a high level of structural variation. Our findings show that while core genomes of the multiple introductions are highly similar, the mini‐chromosomes have undergone marked diversification. The maintenance of the mini‐chromosome and rapid genomic changes suggest the mini‐chromosomes may serve important virulence or niche adaptation roles under diverse environmental conditions.

Plant Sciences↗

Engineering well-expressed, V2-immunofocusing HIV-1 envelope glycoprotein membrane trimers for use in heterologous prime-boost vaccine regimens

HIV-1 vaccine immunofocusing strategies may be able to induce broadly-reactive neutralizing antibodies (NAbs). Here, we engineered a panel of diverse, membrane-resident native HIV-1 trimers vulnerable to two broad targets—the V2 apex and fusion peptide (FP). Selection criteria included i) high expression and ii) infectious function, so that trimer neutralization sensitivity can be profiled in pseudovirus (PV) assays. Initially, we boosted the expression of 17 candidate trimers by truncating gp41 and introducing a gp120-gp41 SOS disulfide to prevent gp120 shedding. "Repairs" were made to fill glycan holes and eliminate other strain-specific aberrations. A new neutralization assay allowed PV infection when our standard assay was insufficient. Trimers with exposed V3 loops, a target of non-NAbs, were discarded. To try to increase V2-sensitivity, we removed clashing glycans and modified the C-strand. Notably, a D167N mutation improved V2-sensitivity in several cases. Glycopeptide analysis of JR-FL trimers revealed near complete sequon occupation and that filling the N197 glycan hole was well-tolerated. In contrast, sequon optimization and inserting/removing glycans at other positions frequently had global "ripple" effects on glycan maturation and sequon occupation throughout the gp120 outer domain and gp41. V2 MAb CH01 selectively bound to trimers with small high mannose glycans near the base of the V1 loop, thereby avoiding clashes. Knocking in a rare N49 glycan was found to perturb gp41 glycans, increasing FP NAb sensitivity—and sometimes improving expression. Finally, a biophysical analysis of VLPs revealed that i) ~25% of particles bear Env spikes, ii) spontaneous particle budding is high and only increases 4-fold upon Gag transfection, and iii) Env+ particles express ~30–40 spikes. Taken together, we identified 7 diverse trimers with a range of sensitivities to two targets to allow rigorous testing of immunofocusing vaccine concepts.

60 APPLIED LIFE SCIENCES↗

Thermal insulation attaching means

An improved isolation system is provided for attaching ceramic tiles of insulating material to the surface of a structure to be protected against extreme temperatures of the nature expected to be encountered by the space shuttle orbiter. This system isolates the fragile ceramic tiles from thermally and mechanically induced vehicle structural strains. The insulating tiles are affixed to a felt isolation pad formed of closely arranged and randomly oriented fibers by means of a flexible adhesive and in turn the felt pad is affixed to the metallic vehicle structure by an additional layer of flexible adhesive.

Leger, L. J.↗